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Enhanced access to the human phosphoproteome with genetically encoded phosphothreonine

Protein phosphorylation is a ubiquitous post-translational modification used to regulate cellular processes and proteome architecture by modulating protein-protein interactions. The identification of phosphorylation events through proteomic surveillance has dramatically outpaced our capacity for fun...

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Autores principales: Moen, Jack M., Mohler, Kyle, Rogulina, Svetlana, Shi, Xiaojian, Shen, Hongying, Rinehart, Jesse
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9700786/
https://www.ncbi.nlm.nih.gov/pubmed/36433969
http://dx.doi.org/10.1038/s41467-022-34980-5
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author Moen, Jack M.
Mohler, Kyle
Rogulina, Svetlana
Shi, Xiaojian
Shen, Hongying
Rinehart, Jesse
author_facet Moen, Jack M.
Mohler, Kyle
Rogulina, Svetlana
Shi, Xiaojian
Shen, Hongying
Rinehart, Jesse
author_sort Moen, Jack M.
collection PubMed
description Protein phosphorylation is a ubiquitous post-translational modification used to regulate cellular processes and proteome architecture by modulating protein-protein interactions. The identification of phosphorylation events through proteomic surveillance has dramatically outpaced our capacity for functional assignment using traditional strategies, which often require knowledge of the upstream kinase a priori. The development of phospho-amino-acid-specific orthogonal translation systems, evolutionarily divergent aminoacyl-tRNA synthetase and tRNA pairs that enable co-translational insertion of a phospho-amino acids, has rapidly improved our ability to assess the physiological function of phosphorylation by providing kinase-independent methods of phosphoprotein production. Despite this utility, broad deployment has been hindered by technical limitations and an inability to reconstruct complex phopho-regulatory networks. Here, we address these challenges by optimizing genetically encoded phosphothreonine translation to characterize phospho-dependent kinase activation mechanisms and, subsequently, develop a multi-level protein interaction platform to directly assess the overlap of kinase and phospho-binding protein substrate networks with phosphosite-level resolution.
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spelling pubmed-97007862022-11-27 Enhanced access to the human phosphoproteome with genetically encoded phosphothreonine Moen, Jack M. Mohler, Kyle Rogulina, Svetlana Shi, Xiaojian Shen, Hongying Rinehart, Jesse Nat Commun Article Protein phosphorylation is a ubiquitous post-translational modification used to regulate cellular processes and proteome architecture by modulating protein-protein interactions. The identification of phosphorylation events through proteomic surveillance has dramatically outpaced our capacity for functional assignment using traditional strategies, which often require knowledge of the upstream kinase a priori. The development of phospho-amino-acid-specific orthogonal translation systems, evolutionarily divergent aminoacyl-tRNA synthetase and tRNA pairs that enable co-translational insertion of a phospho-amino acids, has rapidly improved our ability to assess the physiological function of phosphorylation by providing kinase-independent methods of phosphoprotein production. Despite this utility, broad deployment has been hindered by technical limitations and an inability to reconstruct complex phopho-regulatory networks. Here, we address these challenges by optimizing genetically encoded phosphothreonine translation to characterize phospho-dependent kinase activation mechanisms and, subsequently, develop a multi-level protein interaction platform to directly assess the overlap of kinase and phospho-binding protein substrate networks with phosphosite-level resolution. Nature Publishing Group UK 2022-11-24 /pmc/articles/PMC9700786/ /pubmed/36433969 http://dx.doi.org/10.1038/s41467-022-34980-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Moen, Jack M.
Mohler, Kyle
Rogulina, Svetlana
Shi, Xiaojian
Shen, Hongying
Rinehart, Jesse
Enhanced access to the human phosphoproteome with genetically encoded phosphothreonine
title Enhanced access to the human phosphoproteome with genetically encoded phosphothreonine
title_full Enhanced access to the human phosphoproteome with genetically encoded phosphothreonine
title_fullStr Enhanced access to the human phosphoproteome with genetically encoded phosphothreonine
title_full_unstemmed Enhanced access to the human phosphoproteome with genetically encoded phosphothreonine
title_short Enhanced access to the human phosphoproteome with genetically encoded phosphothreonine
title_sort enhanced access to the human phosphoproteome with genetically encoded phosphothreonine
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9700786/
https://www.ncbi.nlm.nih.gov/pubmed/36433969
http://dx.doi.org/10.1038/s41467-022-34980-5
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